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The objective of the present study was to determine the effects of replacing glucose with pyruvate and lactate during the first 48 h of in vitro culture (IVC) in NCSU-23 medium on embryo development, embryo quality and survival of porcine blastocysts after vitrification. To this end, in vitro-produced (IVP) porcine oocytes were cultured with either glucose for 6 days (IVC-Glu) or pyruvate-lactate from Day 0 to Day 2 and then with glucose until Day 6 (IVC-PyrLac). Blastocysts were vitrified on Day 6 using the Cryotop device and, after warming, survival rate and the apoptosis index were evaluated after 24 h incubation in NCSU-23 medium. No significant differences were observed between IVC-Glu and IVC-PyrLac in terms of cleavage rate, blastocyst yield, total number of cells per blastocyst or the apoptosis index (1.82±0.75% vs 3.18±0.88%, respectively) of non-vitrified embryos. However, a significant increase was seen in hatching/hatched blastocysts in the IVC-PyrLac compared with IVC-Glu treatment group (12.71±1.20% vs 3.54±0.47%, respectively). Regardless of treatment, vitrification impaired the survival rate and the apoptosis index. When comparing both treatments after warming, the percentage of apoptotic cells was significantly higher for blastocysts in the IVC-PyrLac compared with IVC-Glu group (18.55±3.49% vs 9.12±2.17%, respectively). In conclusion, under the conditions of the present study, replacement of glucose with pyruvate-lactate during the first 48 h of culture resulted in a lower cryotolerance of IVP porcine embryos.

Original publication

DOI

10.1071/RD12117

Type

Journal article

Journal

Reprod Fertil Dev

Publication Date

2013

Volume

25

Pages

737 - 745

Keywords

Age Factors, Animals, Apoptosis, Blastocyst, Cryopreservation, Embryo Culture Techniques, Embryonic Development, Fertilization in Vitro, Glucose, In Situ Nick-End Labeling, Lactic Acid, Pyruvic Acid, Survival Analysis, Swine, Vitrification